Tuberc Respir Dis.  2017 Jul;80(3):255-264. 10.4046/trd.2017.80.3.255.

The Role of N-Acetyl Transferases on Isoniazid Resistance from Mycobacterium tuberculosis and Human: An In Silico Approach

• Unissa AN
• Sukumar S
• Hanna LE

• KMID: 2385040
• DOI: http://doi.org/10.4046/trd.2017.80.3.255

Abstract

BACKGROUND
N-acetyl transferase (NAT) inactivates the pro-drug isoniazid (INH) to N-acetyl INH through a process of acetylation, and confers low-level resistance to INH in Mycobacterium tuberculosis (MTB). Similar to NAT of MTB, NAT2 in humans performs the same function of acetylation. Rapid acetylators, may not respond to INH treatment efficiently, and could be a potential risk factor, for the development of INH resistance in humans.
METHODS
To understand the contribution of NAT of MTB and NAT2 of humans in developing INH resistance using in silico approaches, in this study, the wild type (WT) and mutant (MT)-NATs of MTB, and humans, were modeled and docked, with substrates and product (acetyl CoA, INH, and acetyl INH). The MT models were built, using templates 4BGF of MTB, and 2PFR of humans.
RESULTS
On the basis of docking results of MTB-NAT, it can be suggested that in comparison to the WT, binding affinity of MT-G207R, was found to be lower with acetyl CoA, and higher with acetyl-INH and INH. In case of MT-NAT2 from humans, the pattern of score with respect to acetyl CoA and acetyl-INH, was similar to MT-NAT of MTB, but revealed a decrease in INH score.
CONCLUSION
In MTB, MT-NAT revealed high affinity towards acetyl-INH, which can be interpreted as increased formation of acetyl-INH, and therefore, may lead to INH resistance through inactivation of INH. Similarly, in MT-NAT2 (rapid acetylators), acetylation occurs rapidly, serving as a possible risk factor for developing INH resistance in humans.

Keyword

Mycobacterium tuberculosis; Humans; Isoniazid; Transferases

MeSH Terms

Acetyl Coenzyme A
Acetylation
Computer Simulation*
Humans*
Isoniazid*
Mycobacterium tuberculosis*
Mycobacterium*
Risk Factors
Transferases*
Acetyl Coenzyme A
Isoniazid
Transferases

Figure

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  Figure 1 Isoniazid (INH) is inactivated to acetyl INH by N-acetyltransferase (NAT) enzymes. MTB: Mycobacterium tuberculosis.

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  Figure 2 The generated three-dimensional mutant (MT; Arg207) model (pink) of N-acetyl transferase of Mycobacterium tuberculosis from wild type (WT; Gly207) green in color.

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  Figure 3 The generated three-dimensional mutant (MT) model (Arg268) of N-acetyl transferase 2 of human from wild type (WT; Lys268).

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  Figure 4 Superimposition of WT-4BGF (Gly207) and MT (Arg207) model of N-acetyl transferase from Mycobacterium tuberculosis. WT: wild type; MT: mutant.

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  Figure 5 Superimposition of WT-2PFR (Lys268) and MT (Arg268) model of N-acetyl transferase 2 from human. WT: wild type; MT: mutant.

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  Figure 6 Active residues at ligand binding site in WT-MTB-NAT (green) showing docked isoniazid (pink). WT: wild type; MTB: Mycobacterium tuberculosis; NAT: N-acetyl transferase.

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  Figure 7 Active residues at ligand binding site in WT-human-NAT2 showing docked acetyl isoniazid. WT: wild type; NAT2: N-acetyl transferase 2.

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  Figure 8 (A) Superimposition of WT-4BGF docked with acetyl CoA (green) and 2VFC in complex with CoA (blue). (B) Superimposition of WT-4BGF docked with acetyl isoniazid (green) and 3LTW in complex with hydralazine (purple). WT: wild type.

• 

  Figure 9 (A) Superimposition of WT-2PFR docked with acetyl CoA (red) and 2PFR in complex with CoA (light blue). (B) Superimposition of WT-2PFR docked with isoniazid (red) and 1W6F in complex with isoniazid (mustard). WT: wild type.

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  Figure 10 Docking score of acetyl CoA, isoniazid (INH), and acetyl INH with wild type (WT) and mutant (MT) of N-acetyl transferase from Mycobacterium tuberculosis.

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  Figure 11 Docking score of acetyl CoA, isoniazid (INH), and acetyl INH with wild type (WT) and mutant (MT) of N-acetyl transferase 2 from human.

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  Figure 12 Differences in interactions at ligand binding site between wild type (WT) and mutant (MT) proteins of Mycobacterium tuberculosis. (A) Acetyl CoA. (B) Isoniazid. (C) Acetyl isoniazid.

• 

  Figure 13 Differences in interactions at ligand binding site between wild type (WT) and mutant (MT) proteins of human. (A) Acetyl CoA. (B) Isoniazid. (C) Acetyl isoniazid.

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